Description
Useful for assaying lysates, immunoprecipitates or inhibitor screening using the nuclear extract provided.- Includes HeLa nuclear extract, a rich source of HDACs 1 & 2 for use as a positive control or as a source of HDAC activity for screening.
Compatible with class I & IIb HDAC and sirtuins (with addition of NAD+).
The COLOR DE LYS® system (Colorimetric Histone de Acetylase Lysyl Substrate/Developer) is a sensitive and convenient alternative to protocols utilizing radiolabeled, acetylated histones or peptide/HPLC methods for the assay of histone deacetylases. The assay procedure has two steps. First, the COLOR DE LYS® substrate which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (HeLa nuclear or other extract, purified enzyme, bead bound immunocomplex, etc.). Deacetylation of the substrate sensitizes the substrate so that, in the second step, mixing with the COLOR DE LYS® developer causes an increase in yellow color intensity, and absorption at 405 nm.
Note: The COLOR DE LYS® substrate is efficiently deacetylated by HDAC1 and HDAC2, the major contributors to HDAC activity in HeLa nuclear extracts. It is however, a poor substrate for HDAC3 and recombinant human HDAC8 (BIOMOL International, unpublished results). The activities of other HDAC isotypes with the COLOR DE LYS® substrate have yet to be investigated. HDAC3 and HDAC8 do deacetylate the FLUOR DE LYS® substrate, which is the basis for the HDAC fluorometric activity assay kit (Prod. No. BML-AK500)